WO2022151689A1 - Method for extracting duckweed dormant body starch - Google Patents

Method for extracting duckweed dormant body starch Download PDF

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Publication number
WO2022151689A1
WO2022151689A1 PCT/CN2021/107376 CN2021107376W WO2022151689A1 WO 2022151689 A1 WO2022151689 A1 WO 2022151689A1 CN 2021107376 W CN2021107376 W CN 2021107376W WO 2022151689 A1 WO2022151689 A1 WO 2022151689A1
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Prior art keywords
duckweed
dormant
starch
dormant body
decolorized
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PCT/CN2021/107376
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French (fr)
Chinese (zh)
Inventor
洪雁
王旭
周戚薇
金瑜浈
顾正彪
程力
李兆丰
李才明
班宵逢
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江南大学
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Priority to US17/686,528 priority Critical patent/US20220195073A1/en
Publication of WO2022151689A1 publication Critical patent/WO2022151689A1/en

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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B30/00—Preparation of starch, degraded or non-chemically modified starch, amylose, or amylopectin
    • C08B30/02—Preparatory treatment, e.g. crushing of raw materials or steeping process
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A61K8/732—Starch; Amylose; Amylopectin; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A61Q5/02—Preparations for cleaning the hair
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B30/00—Preparation of starch, degraded or non-chemically modified starch, amylose, or amylopectin
    • C08B30/04—Extraction or purification
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B31/00—Preparation of derivatives of starch
    • C08B31/08—Ethers
    • C08B31/12—Ethers having alkyl or cycloalkyl radicals substituted by heteroatoms, e.g. hydroxyalkyl or carboxyalkyl starch
    • C08B31/125—Ethers having alkyl or cycloalkyl radicals substituted by heteroatoms, e.g. hydroxyalkyl or carboxyalkyl starch having a substituent containing at least one nitrogen atom, e.g. cationic starch
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95

Definitions

  • the invention relates to a method for extracting dormant starch of duckweed, belonging to the technical fields of agriculture and food.
  • Duckweed is the smallest angiosperm in the world. Most of the plants are highly degenerated flat membranous fronds, oblong or obovate. Duckweed generally includes 5 genera and 40 species in 5 genera and 40 species. Duckweed is extremely adaptable to the environment and is widely distributed in lakes, ponds and other still waters around the world. Duckweed is the fastest growing flowering plant in the world. It has a fast reproduction speed and can grow nearly exponentially. It can double in 30 hours under optimal conditions, and its biomass accumulation is more than 2 times faster than that of other flowering plants and conventional crops. Depending on the environment, duckweed controls the lateral sac meristem to form thallus or dormant bodies.
  • duckweed When the duckweed grows in the environment of lack of nutrients and low temperature, the duckweed controls the lateral sac meristem to form a dormant body.
  • the basic components of duckweed dormant body are quite different, and the main components are starch and protein.
  • some varieties can efficiently accumulate starch, and theoretically, more than 2 tons of starch can be produced per mu of water surface per year.
  • duckweed has the advantages of high starch content, easy to harvest, no competition for land with grain, and efficient use of space for artificial breeding, and is a valuable new starch resource.
  • duckweed resources has become a research hotspot, mostly focusing on the influence of growth environment (such as light temperature, hormones, nutrients, heavy metals, etc.) on the growth of duckweed, the systematic classification of duckweed and the screening of specific varieties,
  • growth environment such as light temperature, hormones, nutrients, heavy metals, etc.
  • the genetic engineering of duckweed, the overall utilization of duckweed such as sewage treatment, bioreactor, energy utilization, feed utilization, etc.
  • Starch is widely used in medicine, food, cosmetics, feed, oil drilling, metal casting, textile, papermaking and other industries, and it is of great significance to study the starch of duckweed dormant body.
  • the present invention adopts an organic solvent or an organic solution-water mixture to soak and decolorize the duckweed dormant body, and then utilize a disperser to extract the starch of the duckweed dormant body, because the organic solution and water can enter the dormancy
  • Somatic cells have a certain ability to dissolve fat-soluble pigments and water-soluble pigments, and then most of the pigments in the dormant body of duckweed can be removed to reduce the dissemination of starch; at the same time, disperser treatment can not only make starch from floating Pinga dormant somatic cells can be separated, and the cell walls can be linked to each other, which is beneficial to the later centrifugal separation.
  • the first object of the present invention is to provide a kind of method for duckweed dormant decolorization, comprising the steps:
  • the duckweed dormant body is dried and rehydrated to obtain a rehydrated dormant body; then the rehydrated dormant body is soaked in a solvent, washed and filtered to obtain a decolorized duckweed dormant body.
  • the duckweed dormant body needs to be subjected to impurity removal treatment before drying. Specifically, the mature duckweed dormant body that has fallen on the bottom of the water is collected, and the large dormant body in the dormant body is cleaned by rubbing Some root impurities are removed.
  • the drying is at 40-45°C for 1-3 hours; drying methods include blast drying, airflow drying, low-temperature freeze drying, spin flash drying, and variable temperature and pressure differential puffing drying , vacuum rake drying, low temperature thin layer drying, vacuum low temperature liquid continuous drying or rotary vane drying, the final moisture content of the dormant body after drying is less than or equal to 10%.
  • the purpose of drying is to facilitate preservation and transportation; and second, drying will change the permeability of dormant somatic cells, which is convenient for later decolorization and extraction.
  • secondary impurity removal may be performed by using a winnowing device for the purpose of removing remaining root and thallus impurities.
  • the rehydration conditions are that the rehydration material-to-liquid ratio is 1:5-100 in g/mL, the rehydration time is 20-120 min, and the temperature is 25-50°C; It is further preferred that the rehydration material-liquid ratio is 1:20 (w/v), the rehydration time is 20min, and the temperature is 25°C.
  • the solvent includes, but is not limited to, ethanol, acetone, petroleum ether, and dimethyl sulfoxide; the concentration of the solvent is 40-100% (v/v).
  • the material-liquid ratio of the soaking is 1:10-100 in g/mL, the soaking temperature is 25-50°C, and the soaking time is 4-24h.
  • the soaking can be selected from multiple soaking or single soaking.
  • the duckweed dormant body is a dormant body with higher starch content, and varieties include but are not limited to S.polyrhiza ZH0196, L.aequinoctialis 6002, L.aequinoctialis LC33, L.punctata LC06 , S. polyrhiza LC15, Landoltia OT.
  • the varieties of duckweed belong to, but are not limited to, the genus Polygonatum genus, the genus Rhizoma chinensis, the genus Brassica spp.
  • the filtration is to use filter bag filtration; the washing is to use clean water to wash.
  • the method for decolorization comprises the steps of:
  • the duckweed dormant body is harvested, first-time impurity removal, drying, and secondary impurity removal, the duckweed dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-to-liquid ratio is 1:5 ⁇ 100(w/v), the rehydration time is 20 ⁇ 120min, and the rehydration temperature is 20 ⁇ 50°C;
  • the method for depigmenting the duckweed dormant body further comprises:
  • the precipitate obtained after beating, sieving and centrifuging the fresh or dry rehydrated dormant body is soaked in a solvent, and centrifuged to obtain a decolorized dormant body slurry.
  • the sieving described in the method for decolorizing duckweed dormant bodies is to pass through an 80-mesh sieve, and the centrifugation is centrifugation at 3500 rpm for 10 min.
  • the method for decolorizing comprises the steps:
  • Fresh or rehydrated dormant body After the duckweed dormant body is harvested, impurity removal, beating, sieving and centrifugation, fresh duckweed dormant body mud is obtained;
  • the method for decolorization comprises the steps of:
  • the fresh dormant body was beaten, passed through an 80-mesh sieve, and centrifuged at 3500 rpm for 10 min to obtain a fresh dormant body slurry;
  • the fresh dormant body mud is soaked in the ethanol aqueous solution with a concentration of 85%, centrifuged at 3500rpm for 10min after soaking, and washed twice with sufficient water to obtain the decolorized fresh dormant body mud; wherein the material-to-liquid ratio is 1:5 ( w/v), the soaking temperature is 50°C, and the soaking time is 20min.
  • the purpose of the beating is to destroy the cellular structure of the duckweed dormant body, so as to facilitate decolorization and starch extraction.
  • the second object of the present invention is the decolorized duckweed dormant body and the decolorized dormant body slurry prepared by the method of the present invention.
  • the third object of the present invention is to provide a kind of method of extracting duckweed dormant starch, comprising the steps:
  • the decolorized duckweed dormant body and the decolorized dormant body slurry of the present invention are placed in a dispersion liquid, extracted for 5-60 min under the conditions of 25-50 DEG C and a rotating speed of 6000-18000 rpm, filtered, centrifuged, washed and dried to obtain the obtained Duckweed dormant starch.
  • the solid-liquid ratio of the extraction is 1:10-100 in g/mL.
  • the dispersion liquid includes, but is not limited to, water, ethanol, and acetone.
  • one or more of cellulase, hemicellulase, and pectinase can be selectively added to the dispersion to increase the effect of separating starch from the dormant body;
  • the addition amounts of cellulase, hemicellulase and pectinase are respectively 0.5-3.2U/g, 0.6-2.4U/g and 0.2-1.3U/g.
  • the extraction can be repeated multiple times according to the extraction rate and sample purity.
  • the filtration is sieved with a 200-mesh sieve.
  • the centrifugation is centrifugation at 3000-5000 rpm for 10-50 min.
  • the washing is performed with 50-70% ethanol solution, and the number of washings is 1-5 times.
  • the drying is drying at 40°C to 50°C to a degree of moisture ⁇ 10%.
  • duckweed dormant starch after drying, duckweed dormant starch can be obtained by sifting through a 100-mesh sieve.
  • the described method for extracting duckweed dormant starch comprises the steps:
  • the decolorized duckweed dormant body and the decolorized dormant body slurry are placed in the dispersion liquid according to the material-liquid ratio of 1:10-100 (w/v), and then the dormant body-dispersion liquid system is extracted by a dispersing machine, wherein the dispersing machine is used.
  • the rotating speed is 6000 ⁇ 18000rpm, the processing time is 5 ⁇ 60min, and the processing temperature is 25 ⁇ 50°C; then the extracted feed liquid is filtered, the filter residue is backfilled and dispersed for many times, and the filtrate is collected many times; The collected filtrate is subjected to natural sedimentation and centrifugation to obtain crude starch; finally, the crude starch is washed, dried, pulverized and sieved to obtain duckweed dormant starch.
  • the fourth object of the present invention is the duckweed dormant starch obtained by the method of the present invention.
  • the fifth object of the present invention is the application of the duckweed dormant starch of the present invention in the fields of food, medicine, cosmetics, feed, oil drilling, metal casting, textile and paper making.
  • the application in the food field includes, but is not limited to, the application in steamed bread, bread, cake, biscuits, noodles, fried dough sticks, rice noodles, twist, pancakes or compound rice.
  • the application in the food field includes the application in modified starch, and the application includes but is not limited to acid modified starch, oxidized starch, cross-linked starch, esterified starch, ether Application of starch or graft copolymerization starch.
  • the present invention extracts the duckweed dormant starch from the duckweed dormant body, and compared with common starch resources, the duckweed has a high starch content, does not compete with grains, and is easy to realize large-scale, spatialization Breeding, huge output and other advantages, the development and utilization of duckweed starch, a new resource, has great commercial value.
  • the method of the invention can decolorize the dormant body of duckweed, thereby reducing the dyeing of the dormant body starch by pigments, so that the whiteness of the dormant body starch is improved and the color is better.
  • the decolorizing solution used in the method of the present invention is recyclable and has high economy.
  • the dormant starch obtained by the method of the present invention has high purity and extraction rate, the purity reaches more than 93%, and the maximum can reach more than 96.54%, and the extraction rate reaches more than 85%, and the maximum can reach more than 90.03%.
  • the method of the present invention is simple in process, low in equipment cost, easy to operate, and easy to realize industrialization.
  • Starch is widely used in medicine, food, cosmetics, feed, oil drilling, metal casting, textile, papermaking and other industries, and the high-purity dormant starch extracted by the method of the present invention lays the foundation for the next application of dormant starch , expanding the application of dormant starch.
  • Figure 1 is a picture of the decolorization effect of the intact duckweed dormant body.
  • Figure 2 is an optical microscope image of duckweed dormant starch.
  • Figure 3 is the test result of the polarized cross of duckweed dormant starch.
  • Fig. 4 is the test result of the gelatinization characteristic of duckweed resting body starch.
  • Fig. 5 is the test result of the particle size distribution of duckweed dormant starch.
  • Figure 6 is a centrifugation diagram of starch washing of duckweed dormant bodies.
  • Fig. 7 is the morphological structure diagram of the starch in the dormant body of duckweed.
  • Fig. 8 is the test result of the infrared spectrum of the starch of the duckweed resting body.
  • Fig. 9 is the test result of X-ray diffraction of the starch of duckweed resting body.
  • Figure 10 is a flow chart of the method for extracting duckweed dormant starch.
  • the brightness value L* of the sample is measured by a high-precision colorimeter.
  • the instrument is externally calibrated with a standard blackboard and whiteboard. Black is 0 and white is 100.
  • Use No. 1 ziplock bag as a blank weigh 3g of samples (decolorized duckweed dormant body and duckweed dormant starch) in the No. 1 ziplock bag, flatten it, measure three times in parallel, and take the average value.
  • the determination method of starch whiteness refers to "GB/T 22427.6-2008 Determination of starch whiteness”.
  • the determination method of starch purity refers to the acid hydrolysis method in "GB 5009.9-2016 Determination of Starch in Food Safety National Standard for Food”.
  • the extracted duckweed dormant starch was prepared into a 1% (w/w) aqueous solution, dropped on a glass slide, covered with a cover glass, and observed under an optical microscope (natural light and polarized light).
  • starch powder was fixed on the metal sample stage with conductive double-sided tape, and after gold spraying under vacuum, it was observed under a scanning electron microscope with an accelerating voltage of 3.0 kV.
  • a method for extracting duckweed dormant starch comprising the steps:
  • the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:20 (w/v), the rehydration time is 20min, and the temperature is 25°C;
  • the rehydrated dormant body is filtered dry with a filter bag, then soaked in absolute ethanol, and the dormant body is filtered dry with a filter bag after the soaking, and washed twice with sufficient water to obtain the decolorized duckweed dormant body (effect. Figure 1); wherein the soaking material-liquid ratio is 1:30 (w/v), the soaking temperature is 25°C, and the soaking time is 5h;
  • the decolorized dormant body was filtered dry, and placed in a 30% (v/v) ethanol solution of the dispersion liquid according to the ratio of the material to the liquid of 1:25 (w/v);
  • the disperser rotation speed is 13000rpm
  • the extraction time is 10min
  • the extraction temperature is 25°C;
  • the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
  • the obtained duckweed dormant starch is tested for performance, and the test results are as follows:
  • Figure 3 shows the test results of the polarized cross of duckweed dormant starch. It can be seen from Figure 3 that the intersection point of the polarized cross of duckweed dormant starch is not in the center, the cross is thick, and some even have more than one cross, and the starch granules The black lumps in the middle indicate that some of the duckweed starch granules are semi-composite granules, which may imply the existence of fusion phenomenon during the growth of starch granules.
  • Figure 4 is the test result of the gelatinization characteristics of the starch in the dormant body of duckweed. It can be seen from Figure 4 that the gelatinization temperature of the duckweed starch is low and the viscosity is high; ; Duckweed starch return value is small, indicating that its paste cold viscosity is relatively stable, indicating that it is not easy to retrograde.
  • Fig. 5 is the test result of the particle size distribution of the starch in the dormant body of duckweed. It can be seen from Fig. 5 that the particle size of the starch in the dormant body of duckweed is between 5 and 25um.
  • a method for extracting duckweed dormant starch comprising the steps:
  • the mature duckweed dormant body that fell on the bottom of the water was harvested, most of the root impurities in the dormant body were removed by rubbing, dried at 45°C for 2.5 hours, and then used air separation equipment for secondary impurity removal. Remove remaining roots and thallus impurities to obtain dry dormant bodies;
  • the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:40 (w/v), the rehydration time is 30min, and the temperature is 30°C;
  • the rehydrated dormant body is filtered dry with a filter bag, and then soaked in an ethanol aqueous solution with a concentration of 85%. After the soaking, the dormant body is filtered dry with a filter bag, and washed twice with sufficient water to obtain decolorized duckweed.
  • Dormant body the ratio of material to liquid is 1:5 (w/v), the soaking temperature is 50°C, and the soaking time is 1h;
  • the decolorized dormant body was filtered dry, and placed in the dispersion water according to the ratio of material to liquid of 1:25 (w/v); at the same time, cellulase was added to the dispersion solution, and the addition amount was 1.2U/g substrate (dormant body) ;
  • the disperser rotation speed is 8000rpm
  • the extraction time is 50min
  • the extraction temperature is 50°C;
  • the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
  • a method for extracting duckweed dormant starch comprising the steps:
  • the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:10 (w/v), the rehydration time is 60min, and the temperature is 40°C;
  • the rehydrated dormant body was filtered dry with a filter bag, and then soaked in a solution with a ratio of ethanol and acetone of 7:3 (v/v). Wash twice to obtain decolorized duckweed dormant; wherein the ratio of material to liquid is 1:60 (w/v), the soaking temperature is 40°C, and the soaking time is 8h;
  • the decolorized dormant body is filtered dry, and placed in a dispersion (aqueous ethanol solution with a concentration of 50% (v/v)) according to a material-to-liquid ratio of 1:60 (w/v);
  • the disperser rotation speed is 15000rpm
  • the extraction time is 15min
  • the extraction temperature is 40°C;
  • the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
  • the obtained duckweed dormant starch is tested for performance, and the test results are as follows:
  • a method for extracting duckweed dormant starch comprising the steps:
  • the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:20 (w/v), the rehydration time is 30min, and the temperature is 25°C;
  • the rehydrated dormant body is filtered dry with a filter bag, then soaked in absolute ethanol, after the soaking is finished, the dormant body is filtered dry with a filter bag, and washed twice with sufficient water to obtain a decolorized duckweed dormant body; wherein
  • the soaking material-liquid ratio is 1:30 (w/v), the soaking temperature is 25°C, and the soaking time is 5h.
  • the dormant body after the decolorization is filtered dry, and is placed in the dispersion (water) according to the solid-liquid ratio of 1:40 (w/v);
  • the disperser rotation speed is 12000rpm
  • the extraction time is 15min
  • the extraction temperature is 25°C;
  • the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
  • a method for extracting duckweed dormant starch comprising the steps:
  • the fresh dormant body was beaten, passed through an 80-mesh sieve, and centrifuged at 3500 rpm for 10 min to obtain a fresh dormant body slurry;
  • the fresh dormant body mud is soaked in the ethanol aqueous solution with a concentration of 95%, centrifuged at 4000rpm for 8min after the soaking, and washed twice with sufficient water to obtain the decolorized fresh dormant body mud; wherein the material-to-liquid ratio is 1:5 ( w/v), soaking temperature is 50°C, soaking time is 20min;
  • the disperser rotation speed is 8000rpm
  • the extraction time is 50min
  • the extraction temperature is 50°C;
  • the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
  • the duckweed dormant body raw materials and the duckweed dormant body starch obtained after the decolorization obtained in Examples 1 to 5 were tested for performance, and the test results were as follows:
  • Example Brightness L* BaiDu Extraction rate (%) purity(%) 1 98.02 ⁇ 1.25 94.7 ⁇ 0.3 87.67 ⁇ 1.05 96.24 ⁇ 0.38 2 97.59 ⁇ 2.06 94.4 ⁇ 0.6 89.27 ⁇ 0.93 95.98 ⁇ 0.42 3 97.23 ⁇ 2.36 95.2 ⁇ 0.9 90.03 ⁇ 0.85 96.54 ⁇ 1.21 4 98.56 ⁇ 2.15 95.5 ⁇ 0.5 89.68 ⁇ 0.36 96.34 ⁇ 0.98 5 96.68 ⁇ 3.12 94.5 ⁇ 0.8 89.64 ⁇ 0.62 95.96 ⁇ 1.12
  • Example 6 in the patent "A Compound Modified Cationic Starch and Its Preparation Method" (ZL 201811581189.2), the duckweed dormant starch prepared in Example 4 of the present invention was subjected to acidolysis and etherification modification to prepare Acid-hydrolyzed cationic starch, and with reference to the method of Example 11, acid-hydrolyzed cationic starch was used in shampoo.
  • the foaming property of the shampoo was measured with reference to the measurement method of "GB/T 29679-2013 Shampoo, Shampoo Cream", and the combing property test of the shampoo in the patent "A compound modified cationic starch and its preparation method” Methods The combability of shampoo was measured.
  • Example 1 The temperature of decolorization and soaking in Example 1 was adjusted to 45° C., and the others were the same as those in Example 1, to obtain duckweed dormant starch.
  • Example 2 The decolorizing soaking solution in Example 2 was adjusted to be 30% (v/v) acetone solution, and the others were the same as those in Example 2 to obtain duckweed dormant starch.
  • Example 3 The rotating speed of the dispersing machine in Example 3 was adjusted to be 5000 rpm, and the others were consistent with Example 3 to obtain duckweed dormant starch.
  • the duckweed dormant starch obtained in Examples 7 to 9 is carried out performance test, and the test results are as follows in Table 4:
  • Example 1 The steps of drying, rehydration and decolorization in Example 1 were omitted, and the fresh dormant body was directly extracted, and other conditions were consistent with those of Example 1 to obtain duckweed dormant body starch.
  • Adjust the dispersion step in Example 2 (drain the decolorized dormant body, and place it in the dispersion water according to the material-to-liquid ratio of 1:25 (w/v); add cellulase in the dispersion simultaneously, and the addition amount is 1.2U /g substrate (dormant body)) is pulverized and beaten, and the others are consistent with Example 2 to obtain duckweed dormant body starch.
  • Example 3 The drying, rehydration, and decolorization steps in Example 3 were omitted, and the fresh dormant body was subjected to liquid nitrogen quick-freezing treatment. After being frozen in liquid nitrogen for 24 hours, thawed at room temperature and pulverized (passing through a 200-mesh sieve). Other conditions Consistent with Example 3, duckweed dormant starch was obtained.
  • a method for extracting duckweed dormant starch comprising the steps:
  • the dry dormant body is pulverized and passed through an 80-mesh sieve to obtain dormant body powder;
  • the disperser rotation speed is 12000rpm
  • the extraction time is 10min
  • the extraction temperature is 25°C;
  • the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
  • the obtained decolorized duckweed dormant body was tested for performance, and the test results were as follows: the brightness L* was 56.32 ⁇ 2.36, and a* was -12.35 ⁇ 3.22; the obtained duckweed dormant starch was tested for performance, and the test results were as follows: Brightness The L* was 88.35 ⁇ 1.36, the whiteness was 76.3 ⁇ 0.5, the extraction rate was 85.36 ⁇ 1.33%, and the purity reached 77.92 ⁇ 0.63%.

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Abstract

A method for extracting duckweed dormant body starch. The extraction method comprises the following steps: drying and rehydrating a duckweed dormant body to obtain a rehydrated dormant body; then soaking the rehydrated dormant body in a solvent, washing, and filtering to obtain a decolored duckweed dormant body; or pulping a fresh duckweed dormant body, soaking in a solvent, washing, and filtering to obtain decolored duckweed dormant body pulp; and then putting the decolored duckweed dormant body or the decolored duckweed dormant body pulp into a dispersion liquid, extracting for 5-60 min under the conditions that the temperature is 25-50ºC and the rotating speed is 6000-18000 rpm, filtering, centrifuging, washing and drying to obtain the duckweed dormant body starch. The dormant body starch extracted by the method is high in purity and extraction rate, the purity is 93% or above, the highest purity may be 96.54% or above, and the extraction rate is 85% or above.

Description

一种提取浮萍休眠体淀粉的方法A kind of method for extracting duckweed dormant body starch 技术领域technical field
本发明涉及一种提取浮萍休眠体淀粉的方法，属于农业及食品技术领域。The invention relates to a method for extracting dormant starch of duckweed, belonging to the technical fields of agriculture and food.
背景技术Background technique
浮萍是世界上最小的被子植物，植株形态多为高度退化的扁平的膜质叶状体，呈长圆形或倒卵形。浮萍一般包括多根紫萍属、少根紫萍属、扁平无根萍属、绿萍属和芜萍属，共5个属，40个种。浮萍对环境的适应性极强，广泛分布于全球各地的湖泊、池塘和其他静水水域。浮萍是世界上生长最快的开花植物，其繁殖速度快，可近指数型生长，最适条件下30h即可翻倍，比其他开花植物和常规作物的生物量积累快2倍以上。根据环境的不同，浮萍控制侧囊分生组织形成叶状体或休眠体。当浮萍生长在营养匮乏，低温等环境下，浮萍控制侧囊分生组织形成休眠体。根据其品种的不同、生长环境的差异，浮萍休眠体的基本成分存在较大差异，主要成分为淀粉和蛋白质。通过适当的条件进行休眠体的诱导，部分品种能高效积累淀粉，理论上每年每亩水面可生产淀粉2吨以上。综上所述，浮萍具有淀粉含量高、易于采收、不与粮争地、人工养殖可有效利用空间等优点，是极具价值的淀粉新资源。Duckweed is the smallest angiosperm in the world. Most of the plants are highly degenerated flat membranous fronds, oblong or obovate. Duckweed generally includes 5 genera and 40 species in 5 genera and 40 species. Duckweed is extremely adaptable to the environment and is widely distributed in lakes, ponds and other still waters around the world. Duckweed is the fastest growing flowering plant in the world. It has a fast reproduction speed and can grow nearly exponentially. It can double in 30 hours under optimal conditions, and its biomass accumulation is more than 2 times faster than that of other flowering plants and conventional crops. Depending on the environment, duckweed controls the lateral sac meristem to form thallus or dormant bodies. When the duckweed grows in the environment of lack of nutrients and low temperature, the duckweed controls the lateral sac meristem to form a dormant body. According to different varieties and different growth environments, the basic components of duckweed dormant body are quite different, and the main components are starch and protein. Through the induction of dormant bodies under appropriate conditions, some varieties can efficiently accumulate starch, and theoretically, more than 2 tons of starch can be produced per mu of water surface per year. To sum up, duckweed has the advantages of high starch content, easy to harvest, no competition for land with grain, and efficient use of space for artificial breeding, and is a valuable new starch resource.
近年来，对浮萍资源的开发利用成为研究热点，大多集中于生长环境(如光温、激素、营养物质、重金属等)对浮萍生长的影响、浮萍的系统分类和特定品种的筛选、浮萍基因工程、浮萍的整体利用(如污水处理、生物反应器、能源利用、饲料利用等)，对于浮萍中特定成分的研究较少。而淀粉广泛应用于医药、食品、化妆品、饲料、石油钻井、金属铸造、纺织、造纸等行业，进而对浮萍休眠体淀粉进行研究具有重要意义。In recent years, the development and utilization of duckweed resources has become a research hotspot, mostly focusing on the influence of growth environment (such as light temperature, hormones, nutrients, heavy metals, etc.) on the growth of duckweed, the systematic classification of duckweed and the screening of specific varieties, The genetic engineering of duckweed, the overall utilization of duckweed (such as sewage treatment, bioreactor, energy utilization, feed utilization, etc.), there are few studies on specific components in duckweed. Starch is widely used in medicine, food, cosmetics, feed, oil drilling, metal casting, textile, papermaking and other industries, and it is of great significance to study the starch of duckweed dormant body.
发明内容SUMMARY OF THE INVENTION
[技术问题][technical problem]
休眠体淀粉的研究处于起步阶段，尚未有成熟的浮萍休眠体淀粉提取工艺，经检索，相关专利采用了液氮冷冻粉碎提取休眠体淀粉，其成本高，难于工业化；科研论文中报道的提取浮萍休眠体淀粉的方法存在色素浸染淀粉、淀粉提取率低、纯度低、难以工业化等问题，因此，亟需研发一种提取浮萍休眠体淀粉的方法。The research on dormant starch is in its infancy, and there is no mature duckweed dormant starch extraction process. After searching, the relevant patents use liquid nitrogen freezing and crushing to extract dormant starch, which is expensive and difficult to industrialize. The extraction reported in scientific research papers The method of duckweed dormant starch has problems such as pigment-impregnated starch, low starch extraction rate, low purity, and difficulty in industrialization. Therefore, there is an urgent need to develop a method for extracting duckweed dormant starch.
[技术方案][Technical solutions]
为了解决上述至少一个问题，本发明采用有机溶剂或有机溶液-水混合液对浮萍休眠体进行浸泡脱色处理，然后利用分散机对浮萍休眠体淀粉进行提取，由于有机溶液和水能够进入休眠体细胞以及对其中脂溶性色素和水溶性色素的具有一定的溶解能力，进而可以将浮萍休 眠体中大部分色素脱除，减少其对淀粉的浸染；同时分散机处理不仅能使淀粉从浮萍休眠体细胞中分离，又能使细胞壁相互连缀，有益于后期离心分离。In order to solve the above-mentioned at least one problem, the present invention adopts an organic solvent or an organic solution-water mixture to soak and decolorize the duckweed dormant body, and then utilize a disperser to extract the starch of the duckweed dormant body, because the organic solution and water can enter the dormancy Somatic cells have a certain ability to dissolve fat-soluble pigments and water-soluble pigments, and then most of the pigments in the dormant body of duckweed can be removed to reduce the dissemination of starch; at the same time, disperser treatment can not only make starch from floating Pinga dormant somatic cells can be separated, and the cell walls can be linked to each other, which is beneficial to the later centrifugal separation.
本发明的第一个目的是提供一种浮萍休眠体脱色的方法，包括如下步骤：The first object of the present invention is to provide a kind of method for duckweed dormant decolorization, comprising the steps:
将浮萍休眠体干燥、复水，得到复水的休眠体；之后将复水的休眠体在溶剂中浸泡，洗涤、过滤，得到脱色之后的浮萍休眠体。The duckweed dormant body is dried and rehydrated to obtain a rehydrated dormant body; then the rehydrated dormant body is soaked in a solvent, washed and filtered to obtain a decolorized duckweed dormant body.
在本发明的一种实施方式中，浮萍休眠体在干燥之前需要进行除杂处理，具体是将成熟的、掉落在水底的浮萍休眠体采收，经过搓擦将休眠体中的大部分根类杂质去除。In one embodiment of the present invention, the duckweed dormant body needs to be subjected to impurity removal treatment before drying. Specifically, the mature duckweed dormant body that has fallen on the bottom of the water is collected, and the large dormant body in the dormant body is cleaned by rubbing Some root impurities are removed.
在本发明的一种实施方式中，所述的干燥是40～45℃干燥1～3h；干燥的方式包括鼓风干燥、气流干燥、低温冷冻干燥、旋转闪蒸式干燥、变温压差膨化干燥、真空耙式干燥、低温薄层干燥、真空低温液体连续干燥或旋片式干燥，干燥之后休眠体的最终水分含量≤10％。In an embodiment of the present invention, the drying is at 40-45°C for 1-3 hours; drying methods include blast drying, airflow drying, low-temperature freeze drying, spin flash drying, and variable temperature and pressure differential puffing drying , vacuum rake drying, low temperature thin layer drying, vacuum low temperature liquid continuous drying or rotary vane drying, the final moisture content of the dormant body after drying is less than or equal to 10%.
在本发明的一种实施方式中，所述的干燥的目的一是便于保存和运输；二是干燥会改变休眠体细胞的通透性，便于后期脱色和提取。In one embodiment of the present invention, the purpose of drying is to facilitate preservation and transportation; and second, drying will change the permeability of dormant somatic cells, which is convenient for later decolorization and extraction.
在本发明的一种实施方式中，干燥之后还可以利用风选设备进行二次除杂，目的是除去剩余的根类和叶状体杂质。In an embodiment of the present invention, after drying, secondary impurity removal may be performed by using a winnowing device for the purpose of removing remaining root and thallus impurities.
在本发明的一种实施方式中，所述的复水的条件是复水料液比以g/mL计为1：5～100，复水时间为20～120min，温度为25～50℃；进一步优选为复水料液比为1：20(w/v)，复水时间为20min，温度为25℃。In one embodiment of the present invention, the rehydration conditions are that the rehydration material-to-liquid ratio is 1:5-100 in g/mL, the rehydration time is 20-120 min, and the temperature is 25-50°C; It is further preferred that the rehydration material-liquid ratio is 1:20 (w/v), the rehydration time is 20min, and the temperature is 25°C.
在本发明的一种实施方式中，所述的溶剂包括但不限于乙醇、丙酮、石油醚、二甲亚砜；溶剂的浓度为40～100％(v/v)。In an embodiment of the present invention, the solvent includes, but is not limited to, ethanol, acetone, petroleum ether, and dimethyl sulfoxide; the concentration of the solvent is 40-100% (v/v).
在本发明的一种实施方式中，所述的浸泡的料液比以g/mL计为1：10～100，浸泡温度为25～50℃，浸泡时间为4～24h。In an embodiment of the present invention, the material-liquid ratio of the soaking is 1:10-100 in g/mL, the soaking temperature is 25-50°C, and the soaking time is 4-24h.
在本发明的一种实施方式中，所述的浸泡可选择多次浸泡或单次浸泡。In an embodiment of the present invention, the soaking can be selected from multiple soaking or single soaking.
在本发明的一种实施方式中，所述的浮萍休眠体为淀粉含量较高的休眠体，品种包括但不限于S.polyrhiza ZH0196、L.aequinoctialis 6002、L.aequinoctialis LC33、L.punctata LC06、S.polyrhiza LC15、Landoltia OT。In one embodiment of the present invention, the duckweed dormant body is a dormant body with higher starch content, and varieties include but are not limited to S.polyrhiza ZH0196, L.aequinoctialis 6002, L.aequinoctialis LC33, L.punctata LC06 , S. polyrhiza LC15, Landoltia OT.
在本发明的一种实施方式中，所述的浮萍的品种归属但不限于多根紫萍属、少根紫萍属、芜萍属、绿萍属、扁平无根萍属。In one embodiment of the present invention, the varieties of duckweed belong to, but are not limited to, the genus Polygonatum genus, the genus Rhizoma chinensis, the genus Brassica spp.
在本发明的一种实施方式中，所述的过滤是采用滤袋过滤；洗涤是采用清水洗涤。In an embodiment of the present invention, the filtration is to use filter bag filtration; the washing is to use clean water to wash.
在本发明的一种实施方式中，所述脱色的方法包括如下步骤：In one embodiment of the present invention, the method for decolorization comprises the steps of:
(1)浮萍休眠体经采收、一次除杂、干燥、二次除杂后，将浮萍休眠体进行复水，得到 复水的休眠体；其中复水的料液比为1：5～100(w/v)，复水时间为20～120min，复水温度为20～50℃；(1) After the duckweed dormant body is harvested, first-time impurity removal, drying, and secondary impurity removal, the duckweed dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-to-liquid ratio is 1:5 ～100(w/v), the rehydration time is 20～120min, and the rehydration temperature is 20～50℃;
(2)将复水的休眠体滤干，之后放入溶剂中浸泡，洗涤、过滤得到脱色的浮萍休眠体；其中浸泡料液比为1：10(w/v)～1：100(w/v)，浸泡温度为25～50℃，浸泡时间为4～24h。(2) filter the rehydrated dormant body, then put it into a solvent to soak, wash and filter to obtain the decolorized duckweed dormant body; wherein the soaking material-to-liquid ratio is 1:10(w/v)～1:100(w /v), the soaking temperature is 25～50℃, and the soaking time is 4～24h.
在本发明的一种实施方式中，所述的浮萍休眠体脱色的方法还包括：In one embodiment of the present invention, the method for depigmenting the duckweed dormant body further comprises:
新鲜或干燥复水的休眠体打浆、过筛、离心后得到的沉淀物在溶剂中进行浸泡，离心后得到脱色休眠体泥浆。The precipitate obtained after beating, sieving and centrifuging the fresh or dry rehydrated dormant body is soaked in a solvent, and centrifuged to obtain a decolorized dormant body slurry.
在本发明的一种实施方式中，所述的浮萍休眠体脱色的方法中所述的过筛是过80目筛，离心是3500rpm下离心10min。In one embodiment of the present invention, the sieving described in the method for decolorizing duckweed dormant bodies is to pass through an 80-mesh sieve, and the centrifugation is centrifugation at 3500 rpm for 10 min.
在本发明的一种实施方式中，所述脱色的方法包括如下步骤：In one embodiment of the present invention, the method for decolorizing comprises the steps:
(1)新鲜或复水的休眠体：浮萍休眠体经采收、除杂、打浆、过筛、离心后，得到新鲜浮萍休眠体泥浆；(1) Fresh or rehydrated dormant body: After the duckweed dormant body is harvested, impurity removal, beating, sieving and centrifugation, fresh duckweed dormant body mud is obtained;
(2)将新鲜浮萍休眠体泥浆放入溶剂中浸泡，洗涤、离心后得到脱色浮萍休眠体泥浆；其中浸泡料液比为1：10(w/v)～1：100(w/v)，浸泡温度为25～50℃，浸泡时间为4～24h。(2) Put the fresh duckweed dormant body slurry into the solvent to soak, wash and centrifuge to obtain decolorized duckweed dormant body mud; the soaking material-liquid ratio is 1:10(w/v)～1:100(w/v) ), the soaking temperature is 25～50℃, and the soaking time is 4～24h.
在本发明的一种实施方式中，所述脱色的方法包括如下步骤：In one embodiment of the present invention, the method for decolorization comprises the steps of:
将新鲜休眠体进行打浆、过80目筛，3500rpm下离心10min后得到新鲜休眠体泥浆；The fresh dormant body was beaten, passed through an 80-mesh sieve, and centrifuged at 3500 rpm for 10 min to obtain a fresh dormant body slurry;
之后将新鲜休眠体泥浆在浓度为85％的乙醇水溶液中浸泡，浸泡结束之后3500rpm下离心10min，用足量水洗涤两次，得到脱色的新鲜休眠体泥浆；其中料液比为1：5(w/v)，浸泡温度为50℃，浸泡时间为20min。Then the fresh dormant body mud is soaked in the ethanol aqueous solution with a concentration of 85%, centrifuged at 3500rpm for 10min after soaking, and washed twice with sufficient water to obtain the decolorized fresh dormant body mud; wherein the material-to-liquid ratio is 1:5 ( w/v), the soaking temperature is 50°C, and the soaking time is 20min.
在本发明的一种实施方式中，所述的打浆的目的是破坏浮萍休眠体的细胞结构，便于脱色和淀粉提取。In one embodiment of the present invention, the purpose of the beating is to destroy the cellular structure of the duckweed dormant body, so as to facilitate decolorization and starch extraction.
本发明的第二个目的是本发明所述的方法制备得到的脱色的浮萍休眠体和脱色休眠体泥浆。The second object of the present invention is the decolorized duckweed dormant body and the decolorized dormant body slurry prepared by the method of the present invention.
本发明的第三个目的是提供一种提取浮萍休眠体淀粉的方法，包括如下步骤：The third object of the present invention is to provide a kind of method of extracting duckweed dormant starch, comprising the steps:
将本发明所述的脱色的浮萍休眠体和脱色休眠体泥浆置于分散液中，在25～50℃、转速为6000～18000rpm的条件下提取5～60min，过滤、离心、洗涤、干燥得到浮萍休眠体淀粉。The decolorized duckweed dormant body and the decolorized dormant body slurry of the present invention are placed in a dispersion liquid, extracted for 5-60 min under the conditions of 25-50 DEG C and a rotating speed of 6000-18000 rpm, filtered, centrifuged, washed and dried to obtain the obtained Duckweed dormant starch.
在本发明的一种实施方式中，所述提取的料液比以g/mL计为1：10～100。In an embodiment of the present invention, the solid-liquid ratio of the extraction is 1:10-100 in g/mL.
在本发明的一种实施方式中，所述分散液包括但不限于水、乙醇、丙酮。In an embodiment of the present invention, the dispersion liquid includes, but is not limited to, water, ethanol, and acetone.
在本发明的一种实施方式中，所述分散液中可选择性添加纤维素酶、半纤维素酶、果胶酶中的一种或几种，以增益淀粉从休眠体中分离的效果；其中纤维素酶、半纤维素酶、果胶 酶的添加量分别为0.5～3.2U/g、0.6～2.4U/g、0.2～1.3U/g。In one embodiment of the present invention, one or more of cellulase, hemicellulase, and pectinase can be selectively added to the dispersion to increase the effect of separating starch from the dormant body; The addition amounts of cellulase, hemicellulase and pectinase are respectively 0.5-3.2U/g, 0.6-2.4U/g and 0.2-1.3U/g.
在本发明的一种实施方式中，所述提取可以根据提取率和样品纯度多次重复提取。In one embodiment of the present invention, the extraction can be repeated multiple times according to the extraction rate and sample purity.
在本发明的一种实施方式中，所述过滤是用200目筛进行筛分。In one embodiment of the present invention, the filtration is sieved with a 200-mesh sieve.
在本发明的一种实施方式中，所述离心是在3000～5000rpm下离心10～50min。In an embodiment of the present invention, the centrifugation is centrifugation at 3000-5000 rpm for 10-50 min.
在本发明的一种实施方式中，所述洗涤是采用50～70％的乙醇溶液进行洗涤，洗涤的次数为1～5次。In an embodiment of the present invention, the washing is performed with 50-70% ethanol solution, and the number of washings is 1-5 times.
在本发明的一种实施方式中，所述干燥是在40℃～50℃下干燥至水分≤10％。In an embodiment of the present invention, the drying is drying at 40°C to 50°C to a degree of moisture≤10%.
在本发明的一种实施方式中，干燥之后可以通过粉粹过100目筛得到浮萍休眠体淀粉。In one embodiment of the present invention, after drying, duckweed dormant starch can be obtained by sifting through a 100-mesh sieve.
在本发明的一种实施方式中，所述的提取浮萍休眠体淀粉的方法，包括如下步骤：In one embodiment of the present invention, the described method for extracting duckweed dormant starch comprises the steps:
将脱色的浮萍休眠体和脱色休眠体泥浆按照料液比为1：10～100(w/v)置于分散液中，之后采用分散机对休眠体-分散液体系进行提取，其中分散机转速为6000～18000rpm，处理时间为5～60min，处理温度为25～50℃；然后对提取后的料液进行过滤，将滤渣进行多次回填、分散，多次收集过滤液；再对多次收集得到的过滤液进行自然沉降、离心，得到粗淀粉；最后对粗淀粉进行洗涤、干燥、磨粉和过筛得到浮萍休眠体淀粉。The decolorized duckweed dormant body and the decolorized dormant body slurry are placed in the dispersion liquid according to the material-liquid ratio of 1:10-100 (w/v), and then the dormant body-dispersion liquid system is extracted by a dispersing machine, wherein the dispersing machine is used. The rotating speed is 6000～18000rpm, the processing time is 5～60min, and the processing temperature is 25～50℃; then the extracted feed liquid is filtered, the filter residue is backfilled and dispersed for many times, and the filtrate is collected many times; The collected filtrate is subjected to natural sedimentation and centrifugation to obtain crude starch; finally, the crude starch is washed, dried, pulverized and sieved to obtain duckweed dormant starch.
本发明的第四个目的是本发明所述的方法提取得到的浮萍休眠体淀粉。The fourth object of the present invention is the duckweed dormant starch obtained by the method of the present invention.
本发明的第五个目的是本发明所述的浮萍休眠体淀粉在食品、医药、化妆品、饲料、石油钻井、金属铸造、纺织、造纸领域的应用。The fifth object of the present invention is the application of the duckweed dormant starch of the present invention in the fields of food, medicine, cosmetics, feed, oil drilling, metal casting, textile and paper making.
在本发明的一种实施方式中，所述在食品领域的应用包括但不限于馒头、面包、蛋糕、饼干、面条、油条、米粉、麻花、煎饼或复配米中的应用。In one embodiment of the present invention, the application in the food field includes, but is not limited to, the application in steamed bread, bread, cake, biscuits, noodles, fried dough sticks, rice noodles, twist, pancakes or compound rice.
在本发明的一种实施方式中，所述在食品领域的应用包括在改性淀粉中的应用，所述的应用包括但不限于酸变性淀粉、氧化淀粉、交联淀粉、酯化淀粉、醚化淀粉或接枝共聚淀粉中的应用。In an embodiment of the present invention, the application in the food field includes the application in modified starch, and the application includes but is not limited to acid modified starch, oxidized starch, cross-linked starch, esterified starch, ether Application of starch or graft copolymerization starch.
[有益效果][Beneficial effect]
(1)本发明从浮萍休眠体中提取得到了浮萍休眠体淀粉，且与常见的淀粉资源相比，浮萍具有淀粉含量高、不与粮争地，易于能实现规模化、空间化养殖，产量巨大等优点，对浮萍淀粉这种新资源的开发利用具有极大的商业价值。(1) the present invention extracts the duckweed dormant starch from the duckweed dormant body, and compared with common starch resources, the duckweed has a high starch content, does not compete with grains, and is easy to realize large-scale, spatialization Breeding, huge output and other advantages, the development and utilization of duckweed starch, a new resource, has great commercial value.
(2)本发明的方法能够使浮萍休眠体脱色，从而减少色素对休眠体淀粉的浸染，使休眠体淀粉白度提高，色泽更好。(2) The method of the invention can decolorize the dormant body of duckweed, thereby reducing the dyeing of the dormant body starch by pigments, so that the whiteness of the dormant body starch is improved and the color is better.
(3)本发明的方法使用的脱色液可回收，经济性高。(3) The decolorizing solution used in the method of the present invention is recyclable and has high economy.
(4)本发明的方法提取得到的休眠体淀粉纯度、提取率较高，纯度达到93％以上，最高 可达96.54％以上，提取率达到85％以上，最高可达90.03％以上。(4) The dormant starch obtained by the method of the present invention has high purity and extraction rate, the purity reaches more than 93%, and the maximum can reach more than 96.54%, and the extraction rate reaches more than 85%, and the maximum can reach more than 90.03%.
(5)本发明的方法工艺简单，设备成本低，易操作，易于实现工业化。(5) The method of the present invention is simple in process, low in equipment cost, easy to operate, and easy to realize industrialization.
(6)淀粉广泛应用于医药、食品、化妆品、饲料、石油钻井、金属铸造、纺织、造纸等行业，而本发明方法提取到的高纯度休眠体淀粉为休眠体淀粉的下一步应用奠定了基础，拓展了休眠体淀粉的应用。(6) Starch is widely used in medicine, food, cosmetics, feed, oil drilling, metal casting, textile, papermaking and other industries, and the high-purity dormant starch extracted by the method of the present invention lays the foundation for the next application of dormant starch , expanding the application of dormant starch.
附图说明Description of drawings
图1为完整浮萍休眠体脱色效果图。Figure 1 is a picture of the decolorization effect of the intact duckweed dormant body.
图2为浮萍休眠体淀粉光学显微镜图。Figure 2 is an optical microscope image of duckweed dormant starch.
图3为浮萍休眠体淀粉的偏光十字的测试结果。Figure 3 is the test result of the polarized cross of duckweed dormant starch.
图4为浮萍休眠体淀粉的糊化特性的测试结果。Fig. 4 is the test result of the gelatinization characteristic of duckweed resting body starch.
图5为浮萍休眠体淀粉的粒径分布的测试结果。Fig. 5 is the test result of the particle size distribution of duckweed dormant starch.
图6为浮萍休眠体淀粉洗涤离心图。Figure 6 is a centrifugation diagram of starch washing of duckweed dormant bodies.
图7为浮萍休眠体淀粉的形貌结构图。Fig. 7 is the morphological structure diagram of the starch in the dormant body of duckweed.
图8为浮萍休眠体淀粉的红外光谱的测试结果。Fig. 8 is the test result of the infrared spectrum of the starch of the duckweed resting body.
图9为浮萍休眠体淀粉的X射线衍射的测试结果。Fig. 9 is the test result of X-ray diffraction of the starch of duckweed resting body.
图10为提取浮萍休眠体淀粉的方法流程图。Figure 10 is a flow chart of the method for extracting duckweed dormant starch.
具体实施方式Detailed ways
以下对本发明的优选实施例进行说明，应当理解实施例是为了更好地解释本发明，不用于限制本发明。The preferred embodiments of the present invention will be described below, and it should be understood that the embodiments are used to better explain the present invention and are not intended to limit the present invention.
测试方法：testing method:
1、亮度的测定方法1. Measurement method of brightness
采用高精度测色仪测定样品的亮度值L*，仪器用标准黑板、白板外部校准，黑色为0，白色为100。用1号自封袋作空白，称取3g样品(脱色的浮萍休眠体和浮萍休眠体淀粉)于1号自封袋中，铺平，测定3次平行，取平均值。The brightness value L* of the sample is measured by a high-precision colorimeter. The instrument is externally calibrated with a standard blackboard and whiteboard. Black is 0 and white is 100. Use No. 1 ziplock bag as a blank, weigh 3g of samples (decolorized duckweed dormant body and duckweed dormant starch) in the No. 1 ziplock bag, flatten it, measure three times in parallel, and take the average value.
2、白度的测定方法2. Determination method of whiteness
淀粉白度的测定方法参照《GB/T 22427.6-2008淀粉白度测定》。The determination method of starch whiteness refers to "GB/T 22427.6-2008 Determination of starch whiteness".
3、淀粉提取率的计算3. Calculation of starch extraction rate
Figure PCTCN2021107376-appb-000001
Figure PCTCN2021107376-appb-000001
式中，ER为淀粉提取率，％；W1为提取淀粉质量，g；W为原料质量，g；A1为提取 淀粉纯度，％；A为原料淀粉纯度，％。In the formula, ER is the starch extraction rate, %; W1 is the extracted starch quality, g; W is the raw material quality, g; A1 is the extracted starch purity, %; A is the raw starch purity, %.
4、淀粉纯度的测试：4. Test of starch purity:
淀粉纯度的测定方法参照《GB 5009.9-2016食品安全国家标准食品中淀粉的测定》中酸水解法。The determination method of starch purity refers to the acid hydrolysis method in "GB 5009.9-2016 Determination of Starch in Food Safety National Standard for Food".
5、淀粉颗粒的光学显微镜观察5. Optical microscope observation of starch granules
将提取得到的浮萍休眠体淀粉配置成1％(w/w)的水溶液，滴一滴在载玻片上，并用盖玻片封片，置于光学显微镜(自然光和偏光)下观察。The extracted duckweed dormant starch was prepared into a 1% (w/w) aqueous solution, dropped on a glass slide, covered with a cover glass, and observed under an optical microscope (natural light and polarized light).
6、淀粉糊化特性的测定6. Determination of starch gelatinization characteristics
称取一定质量淀粉样品和去离子水放入铝盒中，充分搅拌配制成6％(w/w，以干基计)的25g悬浮液。使用RVA测定淀粉的糊化特性，程序设定依据AACC中的Standard 2。A certain mass of starch samples and deionized water were weighed into an aluminum box, and fully stirred to prepare a 25g suspension of 6% (w/w, on a dry basis). The gelatinization properties of starch were determined using RVA, programmed according to Standard 2 in AACC.
7、淀粉的粒径测定7. Determination of particle size of starch
先将100mg淀粉(以干基计)悬浮于蒸馏水(10mL)中，不间断震荡约10min后缓缓加入粒径分析仪(Mastersizer 2000)，以蒸馏水为分散剂，待光强稳定后开始测定。粒子折射率设定值，粒子吸收指数及分散剂折射率设定值分别为1.5，0及1.33。First, 100 mg of starch (on a dry basis) was suspended in distilled water (10 mL), shaken continuously for about 10 minutes, and then slowly added to a particle size analyzer (Mastersizer 2000), using distilled water as a dispersant, and the measurement was started after the light intensity was stable. The set value of the particle refractive index, the particle absorption index and the set value of the refractive index of the dispersant were 1.5, 0 and 1.33, respectively.
8、淀粉颗粒的扫描电子显微镜观察8. Scanning electron microscope observation of starch granules
将少量淀粉粉末用导电双面胶固定在金属样品台上，在真空下进行喷金以后，置于扫描电子显微镜下观察，加速电压为3.0kV。A small amount of starch powder was fixed on the metal sample stage with conductive double-sided tape, and after gold spraying under vacuum, it was observed under a scanning electron microscope with an accelerating voltage of 3.0 kV.
9、淀粉的傅里叶红外光谱扫描9. Fourier infrared spectroscopy scan of starch
精确称取淀粉样品与溴化钾，以1：60(w/w)的比例混合，取仔细研磨后的混合物70mg压成薄片，用用傅里叶红外光谱仪透射模式进行扫描。扣除空气背景，测量范围400-4000cm -1，分辨率为4cm -1。 Accurately weigh the starch sample and potassium bromide, mix them at a ratio of 1:60 (w/w), take 70 mg of the carefully ground mixture and press them into flakes, which are scanned with a Fourier transform infrared spectrometer in transmission mode. After deducting the air background, the measurement range is 400-4000cm -1 and the resolution is 4cm -1 .
实施例1Example 1
一种提取浮萍休眠体淀粉的方法，包括如下步骤：A method for extracting duckweed dormant starch, comprising the steps:
(1)脱色浮萍休眠体的制备：(1) Preparation of decolorized duckweed dormant:
将成熟的、掉落在水底的浮萍休眠体采收，经过搓擦将休眠体中的大部分根类杂质去除，40℃条件下干燥3h，然后利用风选设备进行二次除杂，除去剩余的根类和叶状体杂质，得到干燥的休眠体；Harvest the mature duckweed dormant body that has fallen on the bottom of the water, remove most of the root impurities in the dormant body by rubbing, dry it at 40°C for 3 hours, and then use wind separation equipment for secondary impurity removal to remove The remaining roots and thallus impurities are obtained as dry dormant bodies;
然后将干燥的休眠体进行复水，得到复水的休眠体；其中复水料液比为1：20(w/v)，复水时间为20min，温度为25℃；Then the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:20 (w/v), the rehydration time is 20min, and the temperature is 25°C;
之后将复水的休眠体用滤袋滤干，再在无水乙醇中浸泡，浸泡结束之后用滤袋将休眠体 滤干，用足量水洗涤两次，得到脱色的浮萍休眠体(效果图如图1)；其中浸泡料液比为1：30(w/v)，浸泡温度为25℃，浸泡时间为5h；Afterwards, the rehydrated dormant body is filtered dry with a filter bag, then soaked in absolute ethanol, and the dormant body is filtered dry with a filter bag after the soaking, and washed twice with sufficient water to obtain the decolorized duckweed dormant body (effect. Figure 1); wherein the soaking material-liquid ratio is 1:30 (w/v), the soaking temperature is 25°C, and the soaking time is 5h;
(2)浮萍休眠体淀粉的提取：(2) Extraction of starch from duckweed dormant body:
将脱色的休眠体滤干，按照料液比为1：25(w/v)置于分散液30％(v/v)乙醇溶液中；The decolorized dormant body was filtered dry, and placed in a 30% (v/v) ethanol solution of the dispersion liquid according to the ratio of the material to the liquid of 1:25 (w/v);
之后采用分散机进行提取，分散机转速为13000rpm，提取时间为10min，提取温度为25℃；Then use a disperser for extraction, the disperser rotation speed is 13000rpm, the extraction time is 10min, and the extraction temperature is 25°C;
提取之后将分散后的溶液用200目筛进行筛分，逐步收集滤液；然后对收集到的滤液进行自然沉降，将上清液和滤渣继续回填进行分散；After the extraction, the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
将所有收集到的滤液在3500rpm下离心10min得到沉淀；之后用50％的乙醇溶液对沉淀进行三次洗涤离心(如图2)，再将离心后的沉淀铺平在40℃鼓风干燥箱中干燥至水分≤10％，最后粉粹过100目筛，得到浮萍休眠体淀粉。All collected filtrates were centrifuged at 3500 rpm for 10 min to obtain a precipitate; then the precipitate was washed and centrifuged three times with 50% ethanol solution (as shown in Figure 2), and then the centrifuged precipitate was spread out and dried in a blast drying oven at 40°C. When the moisture content is less than or equal to 10%, the powder is finally sifted through a 100-mesh sieve to obtain the duckweed dormant starch.
将得到的浮萍休眠体淀粉进行性能测试，测试结果如下：The obtained duckweed dormant starch is tested for performance, and the test results are as follows:
光学显微镜图如图2，从图2可以看出：浮萍淀粉颗粒呈椭球状。The optical microscope picture is shown in Figure 2. It can be seen from Figure 2 that the duckweed starch granules are ellipsoid.
图3为浮萍休眠体淀粉的偏光十字的测试结果，从图3可以看出：浮萍休眠体淀粉的偏光十字交点不在中心位置，十字较粗，有的甚至有多于一个交叉，淀粉颗粒中间呈现团块状黑色，表明浮萍淀粉团粒中有一部分是半复合团粒，这可能暗示在淀粉团粒生长过程中存在融合现象。Figure 3 shows the test results of the polarized cross of duckweed dormant starch. It can be seen from Figure 3 that the intersection point of the polarized cross of duckweed dormant starch is not in the center, the cross is thick, and some even have more than one cross, and the starch granules The black lumps in the middle indicate that some of the duckweed starch granules are semi-composite granules, which may imply the existence of fusion phenomenon during the growth of starch granules.
图4为浮萍休眠体淀粉的糊化特性的测试结果，从图4可以看出：浮萍淀粉糊化温度较低、黏度大；浮萍淀粉降落值较大，说明其淀粉糊热稳定性差；浮萍淀粉回值较小，说明其糊冷黏度较为稳定，说明其不易回生。Figure 4 is the test result of the gelatinization characteristics of the starch in the dormant body of duckweed. It can be seen from Figure 4 that the gelatinization temperature of the duckweed starch is low and the viscosity is high; ; Duckweed starch return value is small, indicating that its paste cold viscosity is relatively stable, indicating that it is not easy to retrograde.
图5为浮萍休眠体淀粉的粒径分布的测试结果，从图5可以看出：浮萍休眠体淀粉粒径在5～25um之间。Fig. 5 is the test result of the particle size distribution of the starch in the dormant body of duckweed. It can be seen from Fig. 5 that the particle size of the starch in the dormant body of duckweed is between 5 and 25um.
实施例2Example 2
一种提取浮萍休眠体淀粉的方法，包括如下步骤：A method for extracting duckweed dormant starch, comprising the steps:
(1)脱色的浮萍休眠体的制备：(1) preparation of decolorized duckweed dormant:
将成熟的、掉落在水底的浮萍休眠体采收，经过搓擦将休眠体中的大部分根类杂质去除，45℃条件下干燥2.5h，然后利用风选设备进行二次除杂，除去剩余的根类和叶状体杂质，得到干燥的休眠体；The mature duckweed dormant body that fell on the bottom of the water was harvested, most of the root impurities in the dormant body were removed by rubbing, dried at 45°C for 2.5 hours, and then used air separation equipment for secondary impurity removal. Remove remaining roots and thallus impurities to obtain dry dormant bodies;
然后将干燥的休眠体进行复水，得到复水的休眠体；其中复水料液比为1：40(w/v)， 复水时间为30min，温度为30℃；Then the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:40 (w/v), the rehydration time is 30min, and the temperature is 30°C;
之后将复水的休眠体用滤袋滤干，再在浓度为85％的乙醇水溶液中浸泡，浸泡结束之后用滤袋将休眠体滤干，用足量水洗涤两次，得到脱色的浮萍休眠体；其中料液比为1：5(w/v)，浸泡温度为50℃，浸泡时间为1h；Then, the rehydrated dormant body is filtered dry with a filter bag, and then soaked in an ethanol aqueous solution with a concentration of 85%. After the soaking, the dormant body is filtered dry with a filter bag, and washed twice with sufficient water to obtain decolorized duckweed. Dormant body; the ratio of material to liquid is 1:5 (w/v), the soaking temperature is 50℃, and the soaking time is 1h;
(2)浮萍休眠体淀粉的提取：(2) Extraction of starch from duckweed dormant body:
将脱色的休眠体滤干，按照料液比为1：25(w/v)置于分散液水中；同时在分散液中添加纤维素酶，添加量为1.2U/g底物(休眠体)；The decolorized dormant body was filtered dry, and placed in the dispersion water according to the ratio of material to liquid of 1:25 (w/v); at the same time, cellulase was added to the dispersion solution, and the addition amount was 1.2U/g substrate (dormant body) ;
之后采用分散机进行提取，分散机转速为8000rpm，提取时间为50min，提取温度为50℃；Then use a disperser to extract, the disperser rotation speed is 8000rpm, the extraction time is 50min, and the extraction temperature is 50°C;
提取之后将分散后的溶液用200目筛进行筛分，逐步收集滤液；然后对收集到的滤液进行自然沉降，将上清液和滤渣继续回填进行分散；After the extraction, the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
将所有收集到的滤液在3000rpm下离心15min得到沉淀；之后用70％的乙醇溶液对沉淀进行三次洗涤离心，再将离心后的沉淀铺平在45℃鼓风干燥箱中干燥至水分≤10％，最后粉粹过100目筛，得到浮萍休眠体淀粉。All collected filtrates were centrifuged at 3000 rpm for 15 min to obtain a precipitate; then the precipitate was washed and centrifuged three times with 70% ethanol solution, and then the centrifuged precipitate was spread and dried in a blast drying oven at 45°C until moisture ≤ 10% , and finally sifted through a 100-mesh sieve to obtain duckweed dormant starch.
实施例3Example 3
一种提取浮萍休眠体淀粉的方法，包括如下步骤：A method for extracting duckweed dormant starch, comprising the steps:
(1)脱色的浮萍休眠体的制备：(1) Preparation of decolorized duckweed dormant:
将成熟的、掉落在水底的浮萍休眠体采收，经过搓擦将休眠体中的大部分根类杂质去除，40℃条件下干燥5h，然后利用风选设备进行二次除杂，除去剩余的根类和叶状体杂质，得到干燥的休眠体；Harvest the mature duckweed dormant body that has fallen on the bottom of the water, remove most of the root impurities in the dormant body by rubbing, dry it at 40°C for 5 hours, and then use the wind separation equipment for secondary impurity removal to remove The remaining roots and thallus impurities are obtained as dry dormant bodies;
然后将干燥的休眠体进行复水，得到复水的休眠体；其中复水料液比为1：10(w/v)，复水时间为60min，温度为40℃；Then the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:10 (w/v), the rehydration time is 60min, and the temperature is 40°C;
之后将复水后的休眠体用滤袋滤干，再在乙醇和丙酮比例为7：3(v/v)的溶液中浸泡，浸泡结束之后用滤袋将休眠体滤干，用足量水洗涤两次，得到脱色的浮萍休眠体；其中料液比为1：60(w/v)，浸泡温度为40℃，浸泡时间为8h；After that, the rehydrated dormant body was filtered dry with a filter bag, and then soaked in a solution with a ratio of ethanol and acetone of 7:3 (v/v). Wash twice to obtain decolorized duckweed dormant; wherein the ratio of material to liquid is 1:60 (w/v), the soaking temperature is 40°C, and the soaking time is 8h;
(2)浮萍休眠体淀粉的提取：(2) Extraction of starch from duckweed dormant body:
将脱色的休眠体滤干，按照料液比为1：60(w/v)置于分散液(浓度为50％(v/v)的乙醇水溶液)中；The decolorized dormant body is filtered dry, and placed in a dispersion (aqueous ethanol solution with a concentration of 50% (v/v)) according to a material-to-liquid ratio of 1:60 (w/v);
之后采用分散机进行提取，分散机转速为15000rpm，提取时间为15min，提取温度为40℃；Then use a disperser for extraction, the disperser rotation speed is 15000rpm, the extraction time is 15min, and the extraction temperature is 40°C;
提取之后将分散后的溶液用200目筛进行筛分，逐步收集滤液；然后对收集到的滤液进行自然沉降，将上清液和滤渣继续回填进行分散；After the extraction, the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
将所有收集到的滤液在4500rpm下离心10min得到沉淀；之后用50％的乙醇溶液对沉淀进行三次洗涤离心，再将离心后的沉淀铺平在40℃鼓风干燥箱中干燥至水分≤8％，最后粉粹过100目筛，得到浮萍休眠体淀粉(如图6)。All collected filtrates were centrifuged at 4500 rpm for 10 min to obtain a precipitate; then the precipitate was washed and centrifuged three times with 50% ethanol solution, and then the centrifuged precipitate was spread and dried in a blast drying oven at 40°C until moisture ≤ 8% , and finally sifted through a 100-mesh sieve to obtain duckweed dormant starch (as shown in Figure 6).
将得到的浮萍休眠体淀粉进行性能测试，测试结果如下：The obtained duckweed dormant starch is tested for performance, and the test results are as follows:
颗粒形貌的观察结果如图7，从图7可以看出，浮萍叶状体淀粉的颗粒形貌与浮萍休眠体淀粉一致，呈现椭圆状。The observation results of particle morphology are shown in Figure 7. It can be seen from Figure 7 that the particle morphology of duckweed thallus starch is consistent with that of duckweed dormant starch, showing an oval shape.
淀粉的傅里叶红外光谱结果如图8，从图8可以看出，所测样品为明显的淀粉红外图谱。The results of the Fourier infrared spectrum of starch are shown in Fig. 8. It can be seen from Fig. 8 that the tested sample is an obvious starch infrared spectrum.
X射线衍射图谱结果如图9，从图9可以看出，浮萍叶状体淀粉为C型淀粉。The results of the X-ray diffraction pattern are shown in Figure 9, and it can be seen from Figure 9 that the starch in the duckweed thallus is C-type starch.
实施例4Example 4
一种提取浮萍休眠体淀粉的方法，包括如下步骤：A method for extracting duckweed dormant starch, comprising the steps:
(1)脱色的浮萍休眠体的制备：(1) preparation of decolorized duckweed dormant:
将成熟的、掉落在水底的浮萍休眠体采收，经过搓擦将休眠体中的大部分根类杂质去除，45℃条件下干燥3h，然后利用风选设备进行二次除杂，除去剩余的根类和叶状体杂质，得到干燥的休眠体；Harvest the mature duckweed dormant body that has fallen on the bottom of the water, remove most of the root impurities in the dormant body by rubbing, dry it at 45°C for 3 hours, and then use the wind separation equipment for secondary impurity removal to remove The remaining roots and thallus impurities are obtained as dry dormant bodies;
然后将干燥的休眠体进行复水，得到复水的休眠体；其中复水料液比为1：20(w/v)，复水时间为30min，温度为25℃；Then the dried dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-liquid ratio is 1:20 (w/v), the rehydration time is 30min, and the temperature is 25°C;
之后将复水的休眠体用滤袋滤干，再在无水乙醇中浸泡，浸泡结束之后用滤袋将休眠体滤干，用足量水洗涤两次，得到脱色的浮萍休眠体；其中浸泡料液比为1：30(w/v)，浸泡温度为25℃，浸泡时间为5h，Then the rehydrated dormant body is filtered dry with a filter bag, then soaked in absolute ethanol, after the soaking is finished, the dormant body is filtered dry with a filter bag, and washed twice with sufficient water to obtain a decolorized duckweed dormant body; wherein The soaking material-liquid ratio is 1:30 (w/v), the soaking temperature is 25°C, and the soaking time is 5h.
(2)浮萍休眠体淀粉的提取：(2) Extraction of starch from duckweed dormant body:
将脱色之后的休眠体滤干，按照料液比为1：40(w/v)置于分散液(水)中，；The dormant body after the decolorization is filtered dry, and is placed in the dispersion (water) according to the solid-liquid ratio of 1:40 (w/v);
之后采用分散机进行提取，分散机转速为12000rpm，提取时间为15min，提取温度为25℃；Then use a disperser for extraction, the disperser rotation speed is 12000rpm, the extraction time is 15min, and the extraction temperature is 25°C;
提取之后将分散后的溶液用200目筛进行筛分，逐步收集滤液；然后对收集到的滤液进行自然沉降，将上清液和滤渣继续回填进行分散；After the extraction, the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
将所有收集到的滤液在3500rpm下离心10min得到沉淀；之后用70％的乙醇溶液对沉淀进行三次洗涤离心，再将离心后的沉淀铺平在45℃鼓风干燥箱中干燥至水分≤8％，最后粉 粹过100目筛，得到浮萍休眠体淀粉。All collected filtrates were centrifuged at 3500 rpm for 10 min to obtain a precipitate; then the precipitate was washed and centrifuged three times with 70% ethanol solution, and then the centrifuged precipitate was spread and dried in a blast drying oven at 45°C until moisture ≤ 8% , and finally sifted through a 100-mesh sieve to obtain duckweed dormant starch.
实施例5Example 5
一种提取浮萍休眠体淀粉的方法，包括如下步骤：A method for extracting duckweed dormant starch, comprising the steps:
(1)脱色的浮萍休眠体的制备：(1) preparation of decolorized duckweed dormant:
将成熟的、掉落在水底的浮萍休眠体采收，经过搓擦将休眠体中的大部分根类杂质去除，Harvest the mature duckweed dormant body that has fallen on the bottom of the water, and remove most of the root impurities in the dormant body by rubbing.
将新鲜休眠体进行打浆、过80目筛，3500rpm下离心10min后得到新鲜休眠体泥浆；The fresh dormant body was beaten, passed through an 80-mesh sieve, and centrifuged at 3500 rpm for 10 min to obtain a fresh dormant body slurry;
之后将新鲜休眠体泥浆在浓度为95％的乙醇水溶液中浸泡，浸泡结束之后4000rpm下离心8min，用足量水洗涤两次，得到脱色的新鲜休眠体泥浆；其中料液比为1：5(w/v)，浸泡温度为50℃，浸泡时间为20min；After that, the fresh dormant body mud is soaked in the ethanol aqueous solution with a concentration of 95%, centrifuged at 4000rpm for 8min after the soaking, and washed twice with sufficient water to obtain the decolorized fresh dormant body mud; wherein the material-to-liquid ratio is 1:5 ( w/v), soaking temperature is 50℃, soaking time is 20min;
(2)浮萍休眠体淀粉的提取：(2) Extraction of starch from duckweed dormant body:
将脱色的新鲜休眠体泥浆滤干，按照料液比为1：25(w/v)置于分散液水中；同时在分散液中添加纤维素酶，添加量为1.2U/g底物(休眠体)；Filter the decolorized fresh dormant body mud, and place it in the dispersion water according to the material-to-liquid ratio of 1:25 (w/v). body);
之后采用分散机进行提取，分散机转速为8000rpm，提取时间为50min，提取温度为50℃；Then use a disperser to extract, the disperser rotation speed is 8000rpm, the extraction time is 50min, and the extraction temperature is 50°C;
提取之后将分散后的溶液用200目筛进行筛分，逐步收集滤液；然后对收集到的滤液进行自然沉降，将上清液和滤渣继续回填进行分散；After the extraction, the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
将所有收集到的滤液在3000rpm下离心15min得到沉淀；之后用70％的乙醇溶液对沉淀进行三次洗涤离心，再将离心后的沉淀铺平在45℃鼓风干燥箱中干燥至水分≤10％，最后粉粹过100目筛，得到浮萍休眠体淀粉。All collected filtrates were centrifuged at 3000 rpm for 15 min to obtain a precipitate; then the precipitate was washed and centrifuged three times with 70% ethanol solution, and then the centrifuged precipitate was flattened and dried in a blast drying oven at 45°C until moisture ≤ 10% , and finally sifted through a 100-mesh sieve to obtain duckweed dormant starch.
将实施例1～5得到的脱色之后的浮萍休眠体原料和浮萍休眠体淀粉进行性能测试，测试结果如下：The duckweed dormant body raw materials and the duckweed dormant body starch obtained after the decolorization obtained in Examples 1 to 5 were tested for performance, and the test results were as follows:
表1实施例1～5的脱色之后的浮萍休眠体的测试结果Test results of duckweed dormant bodies after decolorization of Table 1 Examples 1-5
实施例Example 亮度L*Brightness L* a*a*
11 68.56±1.8068.56±1.80 -1.38±0.49-1.38±0.49
22 66.38±1.6466.38±1.64 0.84±0.190.84±0.19
33 70.32±2.0370.32±2.03 0.94±0.260.94±0.26
44 69.62±2.3869.62±2.38 0.63±0.220.63±0.22
55 72.25±1.3572.25±1.35 0.21±0.030.21±0.03
表2实施例1～5的浮萍休眠体淀粉的测试结果The test result of the duckweed resting body starch of table 2 embodiment 1～5
实施例Example 亮度L*Brightness L* 白度BaiDu 提取率(％)Extraction rate (%) 纯度(％)purity(%)
11 98.02±1.2598.02±1.25 94.7±0.394.7±0.3 87.67±1.0587.67±1.05 96.24±0.3896.24±0.38
22 97.59±2.0697.59±2.06 94.4±0.694.4±0.6 89.27±0.9389.27±0.93 95.98±0.4295.98±0.42
33 97.23±2.3697.23±2.36 95.2±0.995.2±0.9 90.03±0.8590.03±0.85 96.54±1.2196.54±1.21
44 98.56±2.1598.56±2.15 95.5±0.595.5±0.5 89.68±0.3689.68±0.36 96.34±0.9896.34±0.98
55 96.68±3.1296.68±3.12 94.5±0.894.5±0.8 89.64±0.6289.64±0.62 95.96±1.1295.96±1.12
实施例6Example 6
参照专利《一种复合变性阳离子淀粉及其制备方法》(ZL 201811581189.2)中实施例6的方法，将本发明实施例4制备得到的浮萍休眠体淀粉进行酸解和醚化改性，制备得到酸解阳离子淀粉，并参照实施例11的方法，将酸解阳离子淀粉用于洗发水中。Referring to the method of Example 6 in the patent "A Compound Modified Cationic Starch and Its Preparation Method" (ZL 201811581189.2), the duckweed dormant starch prepared in Example 4 of the present invention was subjected to acidolysis and etherification modification to prepare Acid-hydrolyzed cationic starch, and with reference to the method of Example 11, acid-hydrolyzed cationic starch was used in shampoo.
参照《GB/T 29679-2013洗发液，洗发膏》的测定方法对洗发水的起泡性进行测定，参照专利《一种复合变性阳离子淀粉及其制备方法》中洗发水的梳理性测试方法对洗发水的梳理性进行测定。The foaming property of the shampoo was measured with reference to the measurement method of "GB/T 29679-2013 Shampoo, Shampoo Cream", and the combing property test of the shampoo in the patent "A compound modified cationic starch and its preparation method" Methods The combability of shampoo was measured.
结果显示酸解阳离子淀粉制备的洗发水指标达到国标要求或市售洗发水对应指标水平。The results showed that the indicators of shampoo prepared by acid hydrolysis of cationic starch reached the national standard or the corresponding indicators of commercial shampoo.
具体的测试结果如下：The specific test results are as follows:
表3实施例6的测试结果The test result of table 3 embodiment 6
样品sample 泡沫高度(mm)Foam height (mm) 湿梳理功(kJ)Wet combing work (kJ) 干梳理功(kJ)Dry combing work (kJ)
酸解阳离子淀粉洗发水Acid hydrolyzed cationic starch shampoo 118±2118±2 13.15±0.0713.15±0.07 7.32±0.117.32±0.11
国标要求National standard requirements ≥50≥50 —— ——
市售水平Commercial level —— ＜13.25<13.25 ＜7.79<7.79
注：表中的“—”代表没有规定。Note: "—" in the table represents no regulation.
实施例7Example 7
调整实施例1中脱色浸泡的温度至45℃，其他和实施例1保持一致，得到浮萍休眠体淀粉。The temperature of decolorization and soaking in Example 1 was adjusted to 45° C., and the others were the same as those in Example 1, to obtain duckweed dormant starch.
实施例8Example 8
调整实施例2中脱色浸泡液为30％(v/v)丙酮溶液，其他和实施例2保持一致，得到浮萍休眠体淀粉。The decolorizing soaking solution in Example 2 was adjusted to be 30% (v/v) acetone solution, and the others were the same as those in Example 2 to obtain duckweed dormant starch.
实施例9Example 9
调整实施例3中分散机的转速为5000rpm，其他和实施例3保持一致，得到浮萍休眠体淀粉。The rotating speed of the dispersing machine in Example 3 was adjusted to be 5000 rpm, and the others were consistent with Example 3 to obtain duckweed dormant starch.
将实施例7～9得到的浮萍休眠体淀粉进行性能测试，测试结果如下表4：The duckweed dormant starch obtained in Examples 7 to 9 is carried out performance test, and the test results are as follows in Table 4:
表4实施例7～9的浮萍休眠体原料和淀粉的测试结果The test result of the duckweed dormant body raw material and starch of table 4 embodiment 7～9
Figure PCTCN2021107376-appb-000002
Figure PCTCN2021107376-appb-000002
对照例1Comparative Example 1
省略实施例1中干燥、复水、脱色的步骤，直接对新鲜休眠体进行提取，其他条件与实施例1保持一致，得到浮萍休眠体淀粉。The steps of drying, rehydration and decolorization in Example 1 were omitted, and the fresh dormant body was directly extracted, and other conditions were consistent with those of Example 1 to obtain duckweed dormant body starch.
对照例2Comparative Example 2
调整实施例2中分散步骤(将脱色的休眠体滤干，按照料液比为1：25(w/v)置于分散液水中；同时在分散液中添加纤维素酶，添加量为1.2U/g底物(休眠体))为粉碎打浆，其他和实施例2保持一致，得到浮萍休眠体淀粉。Adjust the dispersion step in Example 2 (drain the decolorized dormant body, and place it in the dispersion water according to the material-to-liquid ratio of 1:25 (w/v); add cellulase in the dispersion simultaneously, and the addition amount is 1.2U /g substrate (dormant body)) is pulverized and beaten, and the others are consistent with Example 2 to obtain duckweed dormant body starch.
对照例3Comparative Example 3
省略实施例3中的干燥、复水、脱色步骤，对新鲜休眠体进行液氮速冻处理，液氮下冷冻24h后，于室温下解冻，对其进行粉碎处理(过200目筛)，其他条件与实施例3保持一致，得到浮萍休眠体淀粉。The drying, rehydration, and decolorization steps in Example 3 were omitted, and the fresh dormant body was subjected to liquid nitrogen quick-freezing treatment. After being frozen in liquid nitrogen for 24 hours, thawed at room temperature and pulverized (passing through a 200-mesh sieve). Other conditions Consistent with Example 3, duckweed dormant starch was obtained.
将对照例1～3得到的浮萍休眠体淀粉进行性能测试，测试结果如下表5：The duckweed dormant starch obtained in Comparative Examples 1～3 is carried out performance test, and the test results are as follows in Table 5:
表5对照例1～3的浮萍休眠体淀粉的测试结果The test result of the duckweed resting body starch of table 5 comparative examples 1～3
对照例Control example 亮度L*Brightness L* 白度BaiDu 提取率(％)Extraction rate (%) 纯度(％)purity(%)
11 63.03±2.1163.03±2.11 60.1±1.160.1±1.1 42.47±1.7242.47±1.72 83.39±0.3383.39±0.33
22 89.02±1.1389.02±1.13 72.1±0.972.1±0.9 39.98±0.4939.98±0.49 81.06±0.6781.06±0.67
33 65.10±0.4665.10±0.46 69.5±1.369.5±1.3 34.04±0.7134.04±0.71 76.91±0.6176.91±0.61
对照例4Comparative Example 4
一种提取浮萍休眠体淀粉的方法，包括如下步骤：A method for extracting duckweed dormant starch, comprising the steps:
(1)脱色浮萍休眠体的制备：(1) Preparation of decolorized duckweed dormant:
将成熟的、掉落在水底的浮萍休眠体采收，经过搓擦将休眠体中的大部分根类杂质去除，40℃条件下干燥4h，然后利用风选设备进行二次除杂，除去剩余的根类和叶状体杂质，得到干燥的休眠体；Harvest the mature duckweed dormant body that has fallen on the bottom of the water, remove most of the root impurities in the dormant body by rubbing, dry it at 40 ° C for 4 hours, and then use the wind separation equipment for secondary impurity removal to remove The remaining roots and thallus impurities are obtained as dry dormant bodies;
将干燥休眠体进行粉碎处理，过80目筛，得到休眠体粉末；The dry dormant body is pulverized and passed through an 80-mesh sieve to obtain dormant body powder;
使用无水乙醇浸泡休眠体粉末，浸泡结束之后3500rpm下离心10min得到沉淀，用足量水洗涤两次，得到脱色的浮萍休眠体粉末；其中浸泡料液比为1：50(w/v)，浸泡温度为25℃，浸泡时间为10h；Use absolute ethanol to soak the dormant body powder, centrifuge at 3500rpm for 10min after soaking to obtain precipitation, and wash twice with enough water to obtain decolorized duckweed dormant body powder; wherein the soaking material-liquid ratio is 1:50 (w/v) , the soaking temperature is 25℃, and the soaking time is 10h;
(2)浮萍休眠体淀粉的提取：(2) Extraction of starch from duckweed dormant body:
将脱色的休眠体粉末3500rpm下离心10min，按照料液比为1：35(w/v)置于水中；Centrifuge the decolorized dormant body powder at 3500 rpm for 10 min, and place it in water according to the solid-liquid ratio of 1:35 (w/v);
之后采用分散机进行提取，分散机转速为12000rpm，提取时间为10min，提取温度为25℃；Then use a disperser for extraction, the disperser rotation speed is 12000rpm, the extraction time is 10min, and the extraction temperature is 25°C;
提取之后将分散后的溶液用200目筛进行筛分，逐步收集滤液；然后对收集到的滤液进行自然沉降，将上清液和滤渣继续回填进行分散；After the extraction, the dispersed solution is sieved with a 200-mesh sieve, and the filtrate is gradually collected; then the collected filtrate is naturally settled, and the supernatant and the filter residue are continuously backfilled for dispersion;
将所有收集到的滤液在3500rpm下离心10min得到沉淀；之后用70％的乙醇溶液对沉淀进行三次洗涤离心，再将离心后的沉淀铺平在45℃鼓风干燥箱中干燥至水分≤10％，最后粉粹过100目筛，得到浮萍休眠体淀粉。All collected filtrates were centrifuged at 3500 rpm for 10 min to obtain a precipitate; then the precipitate was washed and centrifuged three times with 70% ethanol solution, and then the centrifuged precipitate was spread and dried in a blast drying oven at 45°C until moisture ≤ 10% , and finally sifted through a 100-mesh sieve to obtain duckweed dormant starch.
将得到的脱色的浮萍休眠体进行性能测试，测试结果如下：亮度L*为56.32±2.36，a*为-12.35±3.22；将得到的浮萍休眠体淀粉进行性能测试，测试结果如下：亮度L*为88.35±1.36，白度为76.3±0.5，提取率为85.36±1.33％，纯度达到77.92±0.63％。The obtained decolorized duckweed dormant body was tested for performance, and the test results were as follows: the brightness L* was 56.32 ± 2.36, and a* was -12.35 ± 3.22; the obtained duckweed dormant starch was tested for performance, and the test results were as follows: Brightness The L* was 88.35±1.36, the whiteness was 76.3±0.5, the extraction rate was 85.36±1.33%, and the purity reached 77.92±0.63%.

Claims (19)

  1. 一种浮萍休眠体脱色的方法，其特征在于，包括如下步骤：A method for decolorizing duckweed dormant body, comprising the steps of:
    将浮萍休眠体干燥、复水，得到复水的休眠体；之后将复水的休眠体在溶剂中浸泡，洗涤、过滤，得到脱色之后的浮萍休眠体。The duckweed dormant body is dried and rehydrated to obtain a rehydrated dormant body; then the rehydrated dormant body is soaked in a solvent, washed and filtered to obtain a decolorized duckweed dormant body.
  2. 根据权利要求1所述的方法，其特征在于，所述的复水的条件是复水料液比以g/mL计为1：5～100，复水时间为20～120min，温度为25～50℃。The method according to claim 1, wherein the rehydration conditions are that the rehydration material-to-liquid ratio in g/mL is 1:5～100, the rehydration time is 20～120min, and the temperature is 25～100 50°C.
  3. 根据权利要求1或2所述的方法，其特征在于，所述的溶剂包括但不限于乙醇、丙酮、石油醚、二甲亚砜；溶剂的浓度为40～100％(v/v)。The method according to claim 1 or 2, wherein the solvent includes but is not limited to ethanol, acetone, petroleum ether, and dimethyl sulfoxide; the concentration of the solvent is 40-100% (v/v).
  4. 根据权利要求1或2所述的方法，其特征在于，所述的浸泡的料液比以g/mL计为1：10～100，浸泡温度为25～50℃，浸泡时间为4～24h。The method according to claim 1 or 2, wherein the material-to-liquid ratio of the soaking is 1:10-100 in g/mL, the soaking temperature is 25-50°C, and the soaking time is 4-24h.
  5. 根据权利要求1或2所述的方法，其特征在于，所述的浮萍休眠体为淀粉含量较高的休眠体，品种包括但不限于S.polyrhiza ZH0196、L.aequinoctialis 6002、L.aequinoctialis LC33、L.punctata LC06、S.polyrhiza LC15、Landoltia OT。The method according to claim 1 or 2, wherein the duckweed dormant body is a dormant body with higher starch content, and varieties include but are not limited to S.polyrhiza ZH0196, L.aequinoctialis 6002, L.aequinoctialis LC33 , L.punctata LC06, S.polyrhiza LC15, Landoltia OT.
  6. 根据权利要求1或2所述的方法，其特征在于，所述脱色的方法包括如下步骤：The method according to claim 1 or 2, wherein the decolorizing method comprises the steps:
    (1)浮萍休眠体经采收、一次除杂、干燥、二次除杂后，将浮萍休眠体进行复水，得到复水的休眠体；其中复水的料液比为1：5～100(w/v)，复水时间为20～120min，复水温度为20～50℃；(1) After the duckweed dormant body is harvested, first-time impurity removal, drying, and secondary impurity removal, the duckweed dormant body is rehydrated to obtain a rehydrated dormant body; wherein the rehydration material-to-liquid ratio is 1:5 ～100(w/v), the rehydration time is 20～120min, and the rehydration temperature is 20～50℃;
    (2)将复水的休眠体滤干，之后放入溶剂中浸泡，洗涤、过滤得到脱色的浮萍休眠体；其中浸泡料液比为1：10(w/v)～1：100(w/v)，浸泡温度为25～50℃，浸泡时间为4～24h。(2) filter the rehydrated dormant body, then put it into a solvent to soak, wash and filter to obtain the decolorized duckweed dormant body; wherein the soaking material-to-liquid ratio is 1:10(w/v)～1:100(w /v), the soaking temperature is 25～50℃, and the soaking time is 4～24h.
  7. 根据权利要求1或2所述的方法，其特征在于，所述的浮萍休眠体脱色的方法还包括：The method according to claim 1 or 2, wherein the method for decolorizing the duckweed dormant body further comprises:
    新鲜或干燥复水的休眠体打浆、过筛、离心后得到的沉淀物在溶剂中进行浸泡，离心后得到脱色休眠体泥浆。The precipitate obtained after beating, sieving and centrifuging the fresh or dry rehydrated dormant body is soaked in a solvent, and centrifuged to obtain a decolorized dormant body slurry.
  8. 根据权利要求7所述的方法，其特征在于，所述脱色的方法包括如下步骤：method according to claim 7, is characterized in that, the method for described decolorization comprises the steps:
    (1)新鲜或复水的休眠体：浮萍休眠体经采收、除杂、打浆、过筛、离心后，得到新鲜浮萍休眠体泥浆；(1) Fresh or rehydrated dormant body: After the duckweed dormant body is harvested, impurity removal, beating, sieving and centrifugation, fresh duckweed dormant body mud is obtained;
    (2)将新鲜浮萍休眠体泥浆放入溶剂中浸泡，洗涤、离心后得到脱色浮萍休眠体泥浆；其中浸泡料液比为1：10(w/v)～1：100(w/v)，浸泡温度为25～50℃，浸泡时间为4～24h。(2) Put the fresh duckweed dormant body slurry into the solvent to soak, wash and centrifuge to obtain decolorized duckweed dormant body mud; the soaking material-liquid ratio is 1:10(w/v)～1:100(w/v) ), the soaking temperature is 25～50℃, and the soaking time is 4～24h.
  9. 权利要求1～6任一项所述的方法制备得到的脱色的浮萍休眠体。The decolorized duckweed dormant body prepared by the method of any one of claims 1 to 6.
  10. 权利要求7或8所述的方法制备得到的脱色休眠体泥浆。The decolorized dormant body mud prepared by the method of claim 7 or 8.
  11. 一种提取浮萍休眠体淀粉的方法，其特征在于，包括如下步骤：A method for extracting duckweed dormant starch, comprising the steps of:
    将权利要求9所述的脱色的浮萍休眠体或权利要求10所述的脱色休眠体泥浆置于分散液中，在25～50℃、转速为6000～18000rpm的条件下提取5～60min，过滤、离心、洗涤、干燥得到浮萍休眠体淀粉。The decolorized duckweed dormant body according to claim 9 or the decolorized dormant body slurry according to claim 10 is placed in a dispersion liquid, extracted for 5 to 60 min at 25 to 50° C. and a rotating speed of 6000 to 18000 rpm, and filtered. , centrifugation, washing and drying to obtain duckweed dormant starch.
  12. 根据权利要求11所述的方法，其特征在于，在所述分散液中添加纤维素酶、半纤维素酶、果胶酶中的一种或几种；其中纤维素酶、半纤维素酶、果胶酶的添加量分别为0.5～3.2U/g、0.6～2.4U/g、0.2～1.3U/g。The method according to claim 11, wherein one or more of cellulase, hemicellulase, and pectinase are added to the dispersion; wherein cellulase, hemicellulase, The addition amount of pectinase is 0.5-3.2U/g, 0.6-2.4U/g, 0.2-1.3U/g respectively.
  13. 根据权利要求11或12所述的方法，其特征在于，所述分散液包括但不限于水、乙醇、丙酮。The method according to claim 11 or 12, wherein the dispersion liquid includes but is not limited to water, ethanol, and acetone.
  14. 根据权利要求11或12所述的方法，其特征在于，所述提取的料液比以g/mL计为1：10～100。The method according to claim 11 or 12, characterized in that, the solid-liquid ratio of the extraction is 1:10-100 in g/mL.
  15. 根据权利要求11或12所述的方法，其特征在于，所述的提取浮萍休眠体淀粉的方法，包括如下步骤：method according to claim 11 or 12, is characterized in that, the described method for extracting duckweed dormant starch, comprises the steps:
    将权利要求9所述的脱色的浮萍休眠体或权利要求10所述的脱色休眠体泥浆按照料液比为1：10～100(w/v)置于分散液中，之后采用分散机对休眠体-分散液体系进行提取，其中分散机的转速为6000～18000rpm，处理时间为5～60min，处理温度为25～50℃；然后对提取后的料液进行过滤，将滤渣进行多次回填、分散，多次收集过滤液；再对多次收集得到的过滤液进行自然沉降、离心，得到粗淀粉；最后对粗淀粉进行洗涤、干燥、磨粉和过筛得到浮萍休眠体淀粉。The decolorized duckweed dormant body of claim 9 or the decolorized dormant body slurry of claim 10 are placed in the dispersion liquid according to the material-liquid ratio of 1:10～100 (w/v), and then a disperser is used to The dormant body-dispersion liquid system is extracted, wherein the rotating speed of the disperser is 6000-18000 rpm, the processing time is 5-60 min, and the processing temperature is 25-50 ℃; then the extracted feed liquid is filtered, and the filter residue is backfilled for many times , disperse, and collect the filtrate for many times; then carry out natural sedimentation and centrifugation on the filtrate collected for many times to obtain crude starch; finally, wash, dry, grind and sieve the crude starch to obtain duckweed dormant starch.
  16. 权利要求11～15任一项所述的方法制备得到的浮萍休眠体淀粉。The duckweed dormant starch prepared by the method of any one of claims 11 to 15.
  17. 权利要求16所述的浮萍休眠体淀粉在食品、医药、化妆品、饲料、石油钻井、金属铸造、纺织、造纸领域的应用。The application of the duckweed dormant starch of claim 16 in the fields of food, medicine, cosmetics, feed, oil drilling, metal casting, textile and paper making.
  18. 根据权利要求17所述的应用，其特征在于，所述在食品领域的应用包括但不限于馒头、面包、蛋糕、饼干、面条、油条、米粉、麻花、煎饼或复配米中的应用。The application according to claim 17, wherein the application in the food field includes but is not limited to the application in steamed bread, bread, cake, biscuits, noodles, fried dough sticks, rice flour, twist, pancake or compound rice.
  19. 根据权利要求17所述的应用，其特征在于，所述在食品领域的应用包括在改性淀粉中的应用，所述的应用包括但不限于酸变性淀粉、氧化淀粉、交联淀粉、酯化淀粉、醚化淀粉或接枝共聚淀粉中的应用。The application according to claim 17, wherein the application in the food field includes the application in modified starch, and the application includes but is not limited to acid modified starch, oxidized starch, cross-linked starch, esterification Application of starch, etherified starch or graft copolymerized starch.
PCT/CN2021/107376 2021-01-15 2021-07-20 Method for extracting duckweed dormant body starch WO2022151689A1 (en)

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CN112794920B (en) * 2021-01-15 2022-01-11 江南大学 Method for extracting duckweed dormant body starch
CN114041409A (en) * 2021-08-20 2022-02-15 天津师范大学 Culture method of high-protein duckweed

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